Autochthonous Lactobacillus paracasei subsp. paracasei strains isolated from Caciocavallo cheese: identification and in-vitro investigation on potential probiotic and functional properties

Abstract

Background: In recent years consumers, due to the growing interest for functional foods, have put particularly attention on products enriched with probiotic lactic acid bacteria for their ability to give human benefit by their assumption. Microorganisms to be considered as probiotic should be able to survive to gastrointestinal juices and colonize the intestinal tract to provide benefits to human health. The aim of the study consisted of the in vitro investigation of novel LAB strains, isolated during cheesemaking and ripening of Caciocavallo cheese, and on their probiotic potential for food application. Methods: In order to select potential probiotic bacteria, LAB isolates were preliminarily screened for their survival to simulated gastro-intestinal transitand assayed for other activities of probiotic and functional interest. Thanks to promising attitude as potential probiotics, four strains were tested to evaluate their adhesion ability on Caco-2 cell lines used as intestinal cell model. Microbiological data were analysed by one-way analysis of variance (ANOVA). Comparisons of means were carried out using post-hoc Tukey’s test. For vitro adhesion experiments, statistical significance was evaluated by Kruskal-Wallis test for equal medians. Results: Twenty-five LAB, presumptively identified as Lactobacillus spp., were isolated from cheese during 60 days of ripening and screened for their survival to simulated gastrointestinal transit. Sixteen isolates that showed a survival rate ≥85% were identifiedas 10 different profiles of Lb. paracasei subsp. paracasei. The 10 strains tolerated high concentration of bile salts by Minimal Inhibitory Concentration and growth rate assays and showed susceptibility or moderate susceptibility against antibiotics of human and veterinary importance, except for the resistanceto the class of aminoglycoside antibiotics. Eight out 10 strains showed in vitro cholesterol-lowering ability, whereas all strains showed antioxidant activity of their cell-free supernatants. Moreover, the four strains with highest survival to simulated gastrointestinal transit showed the ability to adhere to Caco-2 cells. Conclusion: The results suggest that some strains may be effective probiotics to be use as tool to design probiotic dairy products after confirmation probiotic activities in further in vivo studies. Findings of the present study suggest that four strains showed good or strong adherent ability on Caco-2 cell monolayer, that is one of prerequisite that probiotic bacteria must have to perform their functional properties. Keywords: adhesion, antioxidant activity, bile salts, cholesterol assimilation, dairy products, gastrointestinal resistance, probiotic lactobacilli.